Ijetr041284

International Journal of Engineering and Technical Research (IJETR) ISSN: 2321-0869 (O) 2454-4698 (P), Volume-4, Issue-2, February 2016

Bioaugmentation novel methods for Biohydrogen production Aysha Sherieff, A. Swaroopa Rani, Zahoorullah.S.MD  Abstract— Bioaugmentation is the improved treatment by adding actively growing and specialized microbial strains into a system to enhance the ability of the process to respond to any fluctuations or to digest or degrade certain compounds.

Whereas Biohydrogen is the production of hydrogen biologically. It occurs most commonly by bacteria, algae and archaea. Biohydrogen is a potential biofuel now-a-days as the non renewable energy resources are depleting, which is obtained from the cultivation of or from waste organic materials. In this paper bioaugmentation novel methods for bio hydrogen production by photo-fermentation bacteria immobilized on agar gel granule and with two different combined bacterial cell cultures (ratios) giving high yield are discussed.

Index Terms— Biohydrogen, Biofuel

Bacteria,

Bioaugmentation,

Biogas,

I. INTRODUCTION The primary methods of biological hydrogen production are Dark and photo-hydrogen productions. During the dark fermentation process the fermentative bacteria uses organic substances as the energy source and electrons for the purpose of metabolic activities and hydrogen production. In photo-fermentation process, organic substrates are utilized as electron donors and light as source of energy [1, 2]. Dark fermentation hydrogen production with the immobilization technology has been widely studied [3,4]. Under mild conditions, the operations to produce hydrogen from biomass and waste materials are conducted that is why the biological hydrogen production is considered an environmentally friendly process [5, 6].

bacteria were immobilized mainly on carrier such as polyvinyl alcohol [9], polyacrylamide [10], or agar gel [11].

The strain KKU PS5 is able to produce hydrogen from lactate [12].

It was shown that enhancement and stabilization of hydrogen production process can be done by immobilized dark fermentative bacteria. For improving the bio-hydrogen process, there are only few reports about the immobilization of photo-fermentation bacteria [13,14]. When hydrogen is burned as a fuel, water is formed or is used to generate the electricity [15]. Hydrogen is a energy carrier and an valuable alternative fuel. It has high energy content ie., 122 kJ/g which is 2.75 times higher than other hydrocarbon fuels [16].

II. IMMOBILIZED BACTERIA The experiment is focused on the factors affecting the photo-hydrogen production. It was shown that immobilized Rhodopseudomonas faecalis RLD-53 exhibited the highest hydrogen yield of 3.15 mol H2/mol acetate under the following optimal conditions. Inoculum age of 24 h , biomass of 4 mg/ml in agar, agar granule diameter of 2.5 mm, agar concentration of 2%, light intensity of 9000 lux. Immobilized photo-fermentation bacteria will not only enhances the hydrogen production but also increase acid-tolerance capacity even at pH 5.0.The immobilized photo-fermentation bacteria can be applied in the combination of dark and photo-fermentation for hydrogen production with high yield [17].

Table1: Different parameters studied for high hydrogen production by Immobilized bacteria [17].

In order to achieve maximum hydrogen production, bioaugmentation of lactic acid-producing bacteria, Lactobacillus delbrueckii ssp. bulgaricus TISTR 895 is used in photo-fermentation process from glucose by purple non-sulfur photosynthetic bacteria (PNSB), Rhodobacter sphaeroides KKU-PS5.

The strain TISTR 895 is a homo-fermentative lactic acid-producing bacteria which produces lactate as its sole end-product [7,8]. In general, Aysha Shereiff, Assistant Professor, Department of Biotechnology, Joginpally B.R Engineering College, Yenkapally, Moinabad Mandal, R.R. District, Hyderabad-500075, Telangana State, India.

A.Swaroopa Rani, Assistant Professor, Department of Biotechnology, JNTUA College of Engineering, Pulivendula-516390, Kadapa District, Andhra Pradesh, India.

Zahoorullah.S.MD, Associate Professor, Department of Biotechnology, Joginpally B.R Engineering College, Yenkapally, Moinabad Mandal, R.R. District, Hyderabad-500075, Telangana State, India.

84

www.erpublication.org

Comparative study of OLR and HRT in different reactors and substrates for Biogas production III. BACTERIAL CULTURE RATIO

under the optimal conditions ie., the optimal initial pH, light intensity, and Mo concentration obtained from RSM with CCD of 7.92, 8.37 klux, and 0.44 mg/L, respectively. PNSB augmented with LAB produced hydrogen from glucose with a relatively high hydrogen yield 9.65 ± 0.23 mol H 2/mol glucose, i.e., 80 % of the theoretical yield. By using appropriate environmental conditions for the cultivation of dark- and photo-fermentative bacteria, bioaugmentation of PNSB with LAB and immobilized photo-fermentation bacteria, an improvement in hydrogen production can be achieved.

Most of the studies are focused on bioaugmentation of native microorganisms that are capable of producing hydrogen with the dark-fermentative hydrogen producers. The information on bioaugmentation of purple non-sulfur photosynthetic bacteria with lactic acid-producing bacteria is limited and not studied in depth. The addition of the desired microorganisms or Bioaugmentation of specialized microbial strains into the anaerobic digesters can enhance the performance of microbial community and also increase the hydrogen production. As a method to produce hydrogen, bioaugmentation of Rhodobacter sphaeroides KKU-PS5 with Lactobacillus delbrueckii ssp. bulgaricus TISTR 895 was conducted.

ACKNOWLEDGMENT The authors would like to thank Mr. Johny Joseph, Sr. Principal Scientist, BEEC, IICT, Tarnaka, Hyderabad, India.

Dr. Ch. Sasikala, Professor, Centre for Environment, Institute of Science & Technology, JNT University, Hyderabad, India and Dr.V.Hima Bindu, Associate Professor, Centre for Environment, Institute of Science & Technology, JNT University, Hyderabad, India.

Because of the differences in the optimal conditions, metabolic types and nutritional requirements of the two bacteria cultivation in the same bioreactor was still difficult even though well-characterized microorganisms were used in the fermentation system. Evaluation of the physical and chemical factors affecting hydrogen production of PNSB (Purple Non-Sulfur photosynthetic Bacteria) augmented with LAB (Lactic Acid-producing Bacteria) was conducted using a full factorial design followed by RSM (Response Surface Methodology) with CCD (Central Composite Design) [18].

REFERENCES [1]. Keskin T, Abo-Hashesh M, Hallenbeck PC. Photo fermentative hydrogen production from wastes.

Bioresour Technol.

2011;102:8557–68.

[2]. Oh YK, Mohan Raj S, Jung GY, Park S. Current status of the metabolic engineering of microorganisms for biohydrogen production. Bioresour Technol. 2011;102:8357–67.

[3] S. K. S. Patel, H. J. Purohit and V. C. Kalia, “Dark Fer-mentative Hydrogen Production by Defined Mixed Mi-crobial Cultures Immobilized on Ligno-Cellulosic Waste Materials,” International Journal of Hydrogen Energy, Vol. 35, No. 19, 2010, pp. 10674-10681. doi:10.1016/j.ijhydene.2010.03.025 [4] J. H. Jo, D. S. Lee, D. Park and J. M. Park, “Biological Hydrogen Production by Immobilized Cells of Clostrid-ium Tyrobutyricum JM1 Isolated from a Food Waste Treatment Process,” Bioresource Technology, Vol. 35, No. 19, 2008, pp. 6666-6672. doi:10.1016/j.biortech.2007.11.067 [5]. Seifert K, Waligorska M, Laniecki M. Hydrogen generation in photobiological process from dairy wastewater. Int J Hydrog Energy.

2010;35:9624–9.

[6]. Sen B, Suttar RR. Mesophilic fermentative hydrogen production from sago starch-processing wastewater using enriched mixed cultures. Int J Hydrog Energy. 2012;37:155884–97.

[7]. Axelsson L. Lactic acid bacteria: classification and physiology. In: Salminen S, Von Wright A, editors. Lactic acid bacteria: microbiology and functional aspects. 2nd ed. New York: Marcel Dekker, Inc; 1998. p.

1–72.

[8]. Hammes WP, Vogel RF. The genus Lactobacillus. In: Wood BJB, Holzapfel WH, editors. The genera of lactic acid bacteria. London: Chapman and Hall. 1995. p. 19–54.

[9] J. O. Kim, Y. H. Kim, J. Y. Ryu, B. K. Song, I. H. Kim and S. H. Yeom, “Immobilization Methods for Continu-ous Hydrogen Gas Production Biofilm Formation Versus Granulation,” Process Biochemistry, Vol. 40, No. 3-4, 2005, pp. 1331-1337. doi:10.1016/j.procbio.2004.06.008 [10] T. Matsunaga, I, Karube and S. Suzuki, “Some Observa-tions on Immobilized Hydrogen-Producing Bacteria: Be-havior of Hydrogen in Gel Membranes,” Biotechnology Bioengineering, Vol. 22, No. 12, 1980, pp. 2607-2615. doi:10.1002/bit.260221209 [11] H. Yokoi, T. Tokushige, J. Hirose, S. Hayashi and Y. Takasaki, “Hydrogen Production by Immobilized Cells of Aciduric Enterobacter Aerogenes Strain HO-39,” Journal of Fermentation and Bioengineering, Vol. 83, No. 5, 1997, pp.

481-484. doi:10.1016/S0922-338X(97)83006-1 [12]. Laocharoen S, Reungsang A. Isolation, characterization and optimization of photo-hydrogen production conditions by newly isolated Rhodobacter sphaeroides KKU-PS5. Int J Hydrog Energy.

2014;39:10870–82.

[13] S. Sawayama, K. K. Rao and D. Hall, “Immobilization of Rhodobacter capsulatus on Cellulose Beads and Water Treatment Using a

Table2: Hydrogen Production and hydrogen yield for Bioaugmentation [18].

IV. CONCLUSION The two Bioaugmentation novel methods analyzed here can be utilized for high biohydrogen production. For immobilized photo-fermentation bacteria, it could promote hydrogen production and the conversion efficiency of substrate by lengthening the time of hydrogen production. It demonstrated that the granule diameter, inoculant age, agar concentration, light intensity and biomass in agar are key factors affecting photo- fermentation hydrogen production and when they are 2.5 mm, 24h, 2%, 4 mg/ml and 7000-9000 lux, the maximum hydrogen yield reached 3.15 mol-H2/mol acetate.

The augmentation of lactic acid-producing bacteria in the purple non-sulfur photosynthetic bacteria fermentation system is beneficial to hydrogen production. Here lactate is serving as another substrate for hydrogen production.

Cumulative hydrogen production of 3396 ± 66 mL H2/L and Hydrogen Production of 9.1 ± 0.2 mL H2/L h were obtained

85

www.erpublication.org

International Journal of Engineering and Technical Research (IJETR) ISSN: 2321-0869 (O) 2454-4698 (P), Volume-4, Issue-2, February 2016 Photobioreactor,” Journal of Fermen-tation and Bioengineering, Vol. 86, No. 5, 1998, pp. 517-520. doi:10.1016/S0922-338X(98)80164-5 [14] X. Tian, Q. Liao, W. Liu, Y. Z. Wang, X. Zhu, J. Li and H. Wang, “Photo-Hydrogen Production Rate of A PVA-Boric Acid Gel Granule Containing Immobilized Photosynthetic Bacteria Cells,” International Journal of Hydrogen Energy, Vol. 34, No. 11, 2009, pp. 4708-4717. doi:10.1016/j.ijhydene.2009.03.042 [15]. Wu TY, Wen Hay JX, Kong LB, Juan JC, Md. Jahim J. Recent advances in reuse of waste material as substrate to produce biohydrogen by purple non-sulfur (PNS) bacteria. Renew Sust Energ Rev. 2012;16:3117–22.

[16]. Gregory DP. The hydrogen economy. Sci Am. 1973;228:13–21.

[17]. Bing-Feng Liu, Guo-Jun Xie, Wan-Qian Guo, Jie Ding, Nan-Qi Ren, “Optimization of Photo-Hydrogen Production by Immobilized Rhodopseudomonas Faecalis RLD-53”, Natural Resources, 2011, 2, 1-7, doi:10.4236/nr.2011.21001.

[18]. Sucheera Laocharoen, Alissara Reungsang and Pensri Plangklang, “Bioaugmentation of Lactobacillus delbrueckii ssp. bulgaricus TISTR 895 to enhance bio‑ hydrogen production of Rhodobacter sphaeroides KKU‑ PS5”, Biotechnol Biofuels (2015) 8:190, DOI 10.1186/s13068-015-0375-z.

86

www.erpublication.org